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Articles by Sanro Tachibana
Total Records ( 15 ) for Sanro Tachibana
  Takaharu Hara , Tomohiro Arita , Sanro Tachibana and Kazutaka Itoh
  For the production of taxol (paclitaxel) by immobilized cells cultured in a bioreactor, cells prepared from Taxus cuspidata var. nana were entrapped with calcium alginate and the amount of paclitaxel released into F4G4 liquid medium was investigated. Immobilized cell cultures were developed using various calcium alginate gel concentrations (2, 2.5 and 4% w/v) and cell concentrations (3, 5 and 10% w/v). Some of the paclitaxel produced in the immobilized cells was found to be released into F4G4 liquid medium. The amount released depended on the gel and cell concentrations used. The free cells in suspension cultures accumulated paclitaxel in intracellular compartments, while the cells immobilized with the calcium alginate gel released some paclitaxel into the medium. The gel forced some of the paclitaxel produced in the cells to be released into the medium. When the cultures using immobilized cells entrapped at a 3% cell concentration and 2.5% gel concentration with an air-lift bioreactor were conducted for 30 days, the amount of paclitaxel released into the medium increased with time on the addition of an elicitor, chito-heptaose. The release of paclitaxel was enhanced to 2.4 mg L-1 of culture medium after the 30-day-incubation. This is the first report that the release of paclitaxel into the medium by immobilized cell cultures of Taxus sp. was enhanced by the addition of an elicitor. Eight five percent of the paclitaxel produced in the cells was released by the immobilized cell cultures in 30 days. The results obtained in this study, with immobilized T. cuspidata var. nana cells in a 2.5% calcium alginate gel, may pave the way for the production of paclitaxel by immobilized cell cultures in a bioreactor.
  Raden Arthur Ario Lelono and Sanro Tachibana
  Eugenia polyantha leaves are widely used in traditional diabetic treatment and as food additives in Indonesia. Extracts of the leaves showed potential inhibitory effect on alpha glucosidase activity in vitro. This study is an effort to evaluate the antidiabetic properties of E. Polyantha leaves extracts. The methanol-water extracts showed the highest activity (IC50 71 μg mL-1), more than the methanol extracts (92 μg mL-1) and water extracts (73 μg mL-1). The extracts at 100 mg kg-1 BW dose also demonstrated a hypoglycemia effect in the in vivo assay test against alloxan induced diabetic rats. Repeated bioassay-guided fractionation of the methanol-water extracts afforded two active compounds, 4-hydroxy-3-methoxy benzoic acid and 4-hydroxy-3, 5-dimethoxy benzoic acid, with 27 and 35% inhibition of alpha glucosidase activity, respectively.
  Sanro Tachibana , Yukinori Kiyota and Michifusa Koga
  To degrade dioxins in contaminated soil, bioremediation was conducted with two fungi (PL1 and 267) screened from nature. A comparison of the concentration of dioxins (Toxicity equivalent quantity) before and after the bioremediation revealed 20 to 90% of dioxins in the soil to be degraded in 15 and 30 days, respectively. Maximum degradation (90%) was obtained with PL1 after 30 days in the presence of 0.1% surfactant.
  Tony Hadibarata , Sanro Tachibana and Kazutaka Itoh
  Microbial degradation of Phenanthrene with several fungi screened from nature was conducted to select fungi for the bioremediation of Phenanthrene. Thrichoderma sp. S019, a fungus collected from soil, had the highest rate of degradation on the agar medium containing Phenanthrene. Maximal degradation (72%) was obtained when Trichoderma sp. S019 was incubated for 30 days after the addition of 0.1 mM of Phenanthrene to the liquid medium. Furthermore, the degradation of Phenanthrene was affected by the addition of a carbon source, the addition of a nitrogen source and agitation. Also, 1,2-Dioxygenase and 2,3-Dioxygenase were produced by Trichoderma sp. S019 in a liquid medium. These enzymes play an important role in the metabolism of substrates, revealing a high stereoselectivity for initial dioxygenase and enzymatic hydration since the K-region of phenanthrene was the major site of metabolism. Phenanthrene was indeed degraded by Trichoderma sp. S019 because 1-Hydroxy-2-naphthoic acid, Salicyaldehyde, Salicylic acid and Catechol, considered to be the intermediates in the bioremediation of Phenanthrene, were detected among the reaction products.
  Sanro Tachibana , Yukinori Kiyota and Michifusa Koga
  To purify dioxin-contaminated soil by bioremediation with the fungi screened from nature, microbial degradation of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin (2,3,7,8-TCDD) was conducted with two fungi (PL1 and 267) screened from nature. The two fungi degraded 22 to 62% of 2,3,7,8-TCDD. Maximum degradation (62%) was obtained when PL1 was incubated for 30 days at 1 ng. Furthermore, bioremediation of 2,3,7,8-TCDD in soil with PL1 was conducted. The PL1 degraded 27 to 51% of the dioxin at 1 and 10 ng g-1 soil in 15 and 30 days, respectively. Maximum degradation (51%) was obtained when PL1 was incubated for 30 days at 1 ng g-1 soil. In addition, 2,3,7,8-TCDD was indeed degraded by the fungi, because 4,5-Dichlorocatechol considered to be an intermediate was detected among the reaction products.
  Tony Hadibarata , Sanro Tachibana and Kazutaka Itoh
  The degradation of n-eicosane by Trichoderma sp. S019, a fungus collected from soil with extensive degradative ability on an agar medium containing n-eicosane, was demonstrated in liquid medium and in soil. Maximal degradation (77%) was obtained when Trichoderma sp. S019 was incubated for 30 days after the addition of 0.1 mM of n-eicosane to the liquid medium while the highest rate of degradation (40%) was obtained in soil with the addition of 1.5% Trichoderma sp. S019. Furthermore, the degradation of n-eicosane was affected by the addition of a carbon source, the addition of a nitrogen source and agitation. n-Eicosane was indeed degraded by Trichoderma sp. S019 because nonadecanoic acid, n-octadecane, hexadecanoic acid, oleic acid and stearic acid, considered to be the intermediates in the biodegradation of n-eicosane, were detected among the reaction products.
  Bambang Wiyono , Sanro Tachibana and Djaban Tinambunan
  Identification of the Diels-Alder adduct of Abietic Acid (AA) and maleic anhydride (MA) or Fumaric Acid (FA) using a Shimadzu QP 5050A Gas chromatograph Mass spectrometer revealed that AA and MA produced endo-maleopimaric acid (MPA) and endo-maleopimaric acid tricarboxylic acid. A reaction product of abietic acid and fumaric acid generated three peaks identified based on their mass spectra as fumaropimaric acid (FPA), FPA adducts and endo-MPA. To maximize the reaction between AA and MA or FA, molar ratio (AA and MA or FA), reaction time and reaction temperature were investigated. The best Diels-Alder reaction between abietic and maleic anhydride was at 125°C for 1 h with a molar ratio of AA to MA of 1: 2. The best Diels-Alder reaction between AA and FA was at 200°C for 1 h with a molar ratio of AA and FA of 1: 2.
  Bambang Wiyono and Sanro Tachibana
  This research is directed at characterizing purified maleopimaric acid (MPA), looking at the amount of MPA and fumaropimaric acid (FPA) made from a large amount of rosin and maleic anhydride (MA) or fumaric acid (FA) with various molar ratios and sources of rosin and evaluating the properties of fortified rosin sizes made from both MPA and FPA. Results showed that identification of the Diels-Alder adduct of abietic acid and maleic anhydride using Mass spectrometry produced a mixture of endo-maleopimaric acid methyl ester with endo-maleopimaric acid tri methyl ester, as indicated by a fragment ion at m/z = 146 with a molecular weight of 414 and fragment ions at m/z = 121, 187, 316 and 386, denoting an endo-maleopimaric acid methyl ester. A fragment ion at m/z = 146 with a molecular weight of 460 and other fragment ions at m/z = 187, 121, 400 and 428 indicated endo-maleopimaric tri methyl ester. Using a large amount of rosin as a raw material to produce MPA, the equation Y = -0.8475 X2 + 10.448X - 9.7125, at a reaction temperature of 200 °C is still relevant as it denoted that a molar ratio of 1:6.2 (rosin and MA) is the best. However, the equation Y = -0.46X2 + 5.268X - 4.47 did not apply to FPA. Using a large amount of rosin, an increase in the molar ratio led to an increase in FPA products. In terms of free rosin and pH, the maleo-and fumaro-pimaric rosin sizes have met the requirement of Indonesian national standards for paste rosin size. In terms of free alkali property, the maleo- and fumaro-pimaric rosin sizes were better than the free alkali of the commercial forms.
  Raden Arthur Ario Lelono , Sanro Tachibana and Kazutaka Itoh
  Antioxidative activities of three bark extracts (methanol, methanol-water and water) from Eugenia polyantha Wight grown in Indonesia were evaluated using various in vitro assays; 2, 2-diphenyl-1-picrylhydrazyl radical-scavenging, hydrogen peroxide-scavenging and β-carotene bleaching assays. From the assays, Eugenia polyantha bark extracts were found to be potential antioxidative activities. The methanol-water extract showed the highest level of free radical-scavenging activity (Effective dose (ED50) = 0.18 mg mL-1) and protection from β-carotene bleaching (85.7% at 100 μg mL-1). The methanol-water extracts showed the highest total phenolic content (856 mg gallic acid equivalent (GAE)/g and 161 mg catechin equivalent (CE)/g) and total antioxidative capacity (449 mg ascorbic acid equivalent (AAE)/g). Furthermore, it exhibited dose-dependent antioxidative activities. A relationship between total antioxidative capacity and total phenolic content was recognized in the three extracts from Eugenia polyantha bark.
  Nina Artanti , Sanro Tachibana , L.B.S. Kardono and Harmastini Sukiman
  Colletotrichum sp. have potential to act as antidiabetic agent, due to its α-glucosidase inhibitory. Therefore, the objective of present study was to isolate and identify the bioactive compounds responsible for the α-glucosidase inhibitory activity in Colletotrichum sp. TSC13. The methanol extract of TSC13 mycelia, was partitioned with n-hexane, chloroform and ethyl acetate. The n-hexane fraction exhibited the strongest α-glucosidase inhibitory activity. Column chromatography of this fraction resulted in 8 sub-fractions (F1-8). Fraction 3 (F3) which showed 71.4±2.4% inhibition was analysed further. Analysis using GC-MS after methylation of F3 and comparison to spectra databases and confirmation using authentic sample standards showed that F3 had two saturated fatty acid methyl esters, palmitic acid and stearic acid methyl esters and three unsaturated fatty acid methyl esters, oleic acid, linoleic acid and linoleinic acid methyl esters. Unsaturated fatty acids showed higher activity than the saturated fatty acids and the methyl esters form of unsaturated fatty acids showed slightly less active than the free acids. Further analysis using an ethyl acetate extract, it was confirmed that most of the fatty acids were present in the form of free acids. Therefore, it was concluded that the α-glucosidase inhibitor compounds in Colletotrichum sp. TSC13 were unsaturated fatty acids. This is the first report that a Colletotrichum sp. from T. sumatrana has α-glucosidase inhibitory activity.
  Raden Arthur Ario Lelono and Sanro Tachibana
  Eugenia polyantha bark extracts were found to have potential antioxidative activities. This study is an effort to investigate the antioxidative compounds in E. polyantha. In vitro antioxidatve assay were used as guided tools for the isolation of antioxidative compounds. The methanol-water extracts showed the highest level of free radical-scavenging activity (ED50) = 180 μg mL-1 and protection from β-carotene bleaching (8.7 μg mL-1). The methanol-water (1:1) extracts exhibited strong DPPH scavenging activity and protection against beta carotene bleaching and was subjected to repeated silica gel column chromatography. The n-butanol, acetone and ethyl acetate solubles exhibited the highest antioxidative activities, derivatization was conducted to the isolated antioxidative compounds prior to identification. Catechin, gallic acid and rutin were isolated from those solubles as active compounds present in the Eugenia polyantha bark.
  Nina Artanti , Sanro Tachibana and Leonardus B.S. Kardono
  The active α-glucosidase inhibitor compounds in the endophytic fungus Colletotrichum sp. TSC13 were found to be the unsaturated fatty acids (oleic, linoleic and linolenic acids). These compounds have potential as antidiabetic agents. The aim of the present study is to investigate the effects of various media composition on growth (mycelium dry weight) and the fatty acids content (μg mg-1 mycelium DW) of Colletotrichum sp. TSC13 in relation to its α-glucosidase inhibitory activity. For that purpose, the experiments were set up by varying the carbon and nitrogen sources, metal ions and desaturase and fatty acid synthase inhibitors in the media. Colletotrichum sp. TSC13 grown on potato dextrose broth (PDB) was used as control. The α-glucosidase inhibitory activities were (range from 43.9±2.5 to 88.6±5.2%) at 10 μg mL-1. This activity seemed to correlate with the unsaturated fatty acids content of the samples. Different sugars as carbon source experiment showed that xylose gave the highest growth (938.7±141.6 mg). However, the highest fatty acids content was obtained from fructose medium which containing linoleic acid (38.8±4.9 μg mg-1 DW). Soluble starch gave better growth (672.5±62.3 mg) but very low fatty acids content (2.8±0.1 μg mg-1 DW) was obtained. Yeast extract was the best nitrogen source. Fatty acids production was better as compared to beef extract and soytone. This is the first report of various media compositions on fatty acids content in Colletotrichum sp. TSC13 in relation to its α-glucosidase inhibitory activity.
  Agus Sukito and Sanro Tachibana
  Two antioxidant active compounds were isolated from the methanol extract of Camellia sasanqua using various in vitro assays: 1,1-diphenyl-2-picrylhydrazyl (DPPH), β-carotene bleaching and reducing power assays. The ethyl acetate (EtOAc) fraction of the methanol extract had the highest DPPH radical-scavenging activity with an Inhibition Concentration (IC50) value of 18.3±1.63 μg mL-1. Sephadex LH-20 column chromatography was used to separate the EtOAc fraction into eight fractions (F1-F8). Antioxidant activity was significantly higher in fraction 5 with an IC50 value 14.61±0.02 μg mL-1. Fraction 5 was further separated by HPLC preparative with Capcellpak C18 MG followed by the Cosmosil 5C18-AR-II column, using a guided DPPH radical-scavenging assay. The compounds isolated were identified as: Hyperoside (1) and isoquercitrin (2) after recrystallization from ethanol, based on Mass Spectrum (MS) and Nuclear Magnetic Resonance (NMR) analyses. Their DPPH radical-scavenging activities based on the 50% scavenging concentration decreased in the following order: Isoquercitrin (21.6 mM)>hyperoside (27.5 mM). The antioxidant activities of hyperoside and isoquercitrin were 67.52±0.64 and 64.33±0.51%, respectively, in the β-carotene bleaching assay. These compounds were found to have good reducing powers (OD value: 2.5-3.8) at concentrations of 50-140 μg mL-1, using the potassium ferricyanide reduction method. Although, these compounds are well-known, hyperoside (1) was isolated from this herb for the first time.
  Amalia Indah Prihantini , Sanro Tachibana and Kazutaka Itoh
  The antioxidant and α-glucosidase inhibitory activities of the methanolic leaf extracts of some subtropical plants were evaluated in the present study. Antioxidant activity was evaluated based on 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, reducing power, hydrogen peroxide and β-carotene bleaching assays. α-Glucosidase inhibitory activity and enzyme kinetics as well as the total phenolic content of the extracts were also investigated. Elaeocarpus sylvestris extract had the highest activities on all the antioxidant assays performed such as DPPH scavenging activity (IC50 12.7±0.5 μg mL-1), reducing power (491.1±6.3 mg QE g -1 dry extract), hydrogen peroxide (IC50 65.6±0.4 μg mL-1) and β-carotene bleaching assays (IC50 5.1±1.9 μg mL-1). The total phenolic content of the E. sylvestris extract also had the highest values for gallic acid, quercetin and rutin equivalents (353.8±28.6 mg GAE g-1 dry extract; 294.9±24.5 mg QE g-1 dry extract; 663.0±52.3 mg RE g-1 dry extract, respectively). α-Glucosidase inhibition assay revealed that Distylium racemosum had the highest activity with an IC50 value of 22.6±1.9 μg mL-1. The results of the present study revealed the potencies of E. sylvestris, D. racemosum, Acer mono Maxim and Liquidambar styraciflua as alternative sources for antioxidants and α-glucosidase inhibitors.
  Agus Sukito and Sanro Tachibana
  In vitro α-glucosidase inhibitory activity of Ginkgo biloba leaves was investigated. The inhibitory activity of methanol extracts from yellow and green leaves was 13.8 and 40.1 μg mL-1, respectively. Each methanol extract was separated into its respective fraction by solvent-solvent extraction with n-hexane, chloroform, ethyl acetate and n-butanol. The n-hexane fractions (in both methanol extracts from green and yellow leaves) exhibited high α-glucosidase inhibitory activity with IC50 values of 13.6 and 13.4 μg mL-1, respectively. Further fractionation of the n-hexane fractions by silica gel column chromatography gave the most active fraction which was identified as ginkgolic acid (C13:0) and a mixture (C13:0, C15:0, C15:1, C17:1 and C17:2). Ginkgolic acid (C13:0) exhibited the highest α-glucosidase inhibitory activity. This is the first study to successfully isolate ginkgolic acids as α-glucosidase inhibitors.
 
 
 
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